Plant Cell Reports

, Volume 37, Issue 12, pp 1625–1637 | Cite as

The wheat TdRL1 is the functional homolog of the rice RSS1 and promotes plant salt stress tolerance

  • Habib Mahjoubi
  • Yutaka Tamari
  • Shin Takeda
  • Oumaya Bouchabké-Coussa
  • Moez Hanin
  • Etienne Herzog
  • Anne-Catherine Schmit
  • Marie-Edith Chabouté
  • Chantal EbelEmail author
Original Article


Key message

Rice rss1 complementation assays show that wheat TdRL1 and RSS1 are true functional homologs. TdRL1 over-expression in Arabidopsis conferred salt stress tolerance and alleviated ROS accumulation.


Plants have developed highly flexible adaptive responses to their ever-changing environment, which are often mediated by intrinsically disordered proteins (IDP). RICE SALT SENSITIVE 1 and Triticum durum RSS1-Like 1 protein (TdRL1) are both IDPs involved in abiotic stress responses, and possess conserved D and DEN-Boxes known to be required for post-translational degradation by the APC/Ccdc20 cyclosome. To further understand their function, we performed a computational analysis to compare RSS1 and TdRL1 co-expression networks revealing common gene ontologies, among which those related to cell cycle progression and regulation of microtubule (MT) networks were over-represented. When over-expressed in Arabidopsis, TdRL1::GFP was present in dividing cells and more visible in cortical and endodermal cells of the Root Apical Meristem (RAM). Incubation with the proteasome inhibitor MG132 stabilized TdRL1::GFP expression in RAM cells showing a post-translational regulation. Moreover, immuno-cytochemical analyses of transgenic roots showed that TdRL1 was present in the cytoplasm and within the microtubular spindle of mitotic cells, while, in interphasic cells, it was rather restricted to the cytoplasm with a spotty pattern at the nuclear periphery. Interestingly in cells subjected to stress, TdRL1 was partly relocated into the nucleus. Moreover, TdRL1 transgenic lines showed increased germination rates under salt stress conditions as compared to wild type. This enhanced salt stress tolerance was associated to an alleviation of oxidative damage. Finally, when expressed in the rice rss1 mutant, TdRL1 suppressed its dwarf phenotype upon salt stress, confirming that both proteins are true functional homologs required for salt stress tolerance in cereals.


Abiotic stress Salt RSS1 Durum wheat TdRL1 Cellular localization Oxidative stress 


Author contribution statement

HM performed the experiments (expression data, cloning, transient and stable transformation of tobacco and Arabidopsis, and germination assays). YT and ST performed rice rss1 complementation assays. OBC developed the pIJBP2 vector for rice transformation. ACS and EH helped with confocal microscopy and immuno-cytochemistry experiments and analyses, and reviewed the article. CE, MH, and MEC designed the experiments, searched for funding, supervised the work, and wrote the article.


This work has been funded by the Tunisian Higher Ministry of Education and the Centre National de la Recherche Scientifique (CNRS). HM was granted from a French-Tunisian bilateral PHC-Utique program (15G0902/32601ZF). Part of this work was granted by the Swiss National Science Foundation and the ‘Mujeres for Africa’ foundation to CE. Microscopy was carried out at the Strasbourg-Esplanade cellular imaging facilities (CNRS, Université de Strasbourg, Région Alsace, Association de la Recherche sur le Cancer, and Ligue Nationale contre le Cancer).

Compliance with ethical standards

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Supplementary material

299_2018_2333_MOESM1_ESM.xlsx (70 kb)
ESM_1: List of genes co-expressed with RSS1 and the GO-term enrichment (excel) (XLSX 69 KB)
299_2018_2333_MOESM2_ESM.xlsx (78 kb)
ESM_ 2: List of genes co-expressed with RSS1 and the GO-term enrichment (excel) (XLSX 78 KB)
299_2018_2333_MOESM3_ESM.pptx (428 kb)
ESM_3: GO-term enrichment in RSS1 co-expressed genes. Illustration done by agriGO (PPTX 428 KB)
299_2018_2333_MOESM4_ESM.pptx (321 kb)
ESM_ 4: GO-term enrichment in TdRL1 co-expressed genes. Illustration done by agriGO (PPTX 321 KB)
299_2018_2333_MOESM5_ESM.xlsx (12 kb)
ESM_5: Gene IDs of genes co-expressed with wheat RL1 and rice RSS1, and used to construct the heatmap in Figure 1b, c. (XLSX 12 KB)
299_2018_2333_MOESM6_ESM.pptx (47 kb)
ESM_ 6: Expression levels of TdRL1 in the different Arabidopsis transgenic lines (PPTX 46 KB)
299_2018_2333_MOESM7_ESM.pptx (389 kb)
ESM_7: Expression of TdRL1-MBD fusion detected by the anti-TdRL1 antibody (PPTX 388 KB)
299_2018_2333_MOESM8_ESM.pptx (309 kb)
ESM_8: Molecular analyses of rss1 complementation lines. (a) Amplification of TdRL1 transgene in rss1 complementation lines, WT and rss1 mutant. (b) Genomic organization of RSS1 WT and rss1 mutant alleles with the position of the Tos17 insertion and of the primers used for genotyping. (c) PCR genotyping of rss1 complemented lines. (PPTX 309 KB)


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  2. Bartels D, Sunkar R (2005) Drought and salt tolerance in plants. Crit Rev Plant Sci 24:23–58CrossRefGoogle Scholar
  3. Batzenschlager M, Lermontova I, Schubert V et al (2015) Arabidopsis MZT1 homologs GIP1 and GIP2 are essential for centromere architecture. PNAS 112:8656–8660. CrossRefPubMedGoogle Scholar
  4. Busso D, Delagoutte-Busso B, Moras D (2005) Construction of a set Gateway-based destination vectors for high-throughput cloning and expression screening in Escherichia coli. Anal Biochem 343:313–321CrossRefGoogle Scholar
  5. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefGoogle Scholar
  6. Cromer L, Jolivet S, Horlow C, Chelysheva L et al (2013) Centromeric cohesion is protected twice at meiosis, by SHUGOSHINs at anaphase I and by PATRONUS at interkinesis. Curr Biol 23:2090–2099CrossRefGoogle Scholar
  7. Ebel C, Hanin M (2016) Maintenance of meristem activity under stress: is there an interplay of RSS1-like proteins with the RBR pathway? Plant Biol 18:167–170. CrossRefPubMedGoogle Scholar
  8. Haak DC, Fukao T, Grene R et al (2017) Multilevel regulation of abiotic stress responses in plants. Front Plant Sci 8:1564. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Hasan MM, Brocca S, Sacco E et al (2014) A comparative study of Whi5 and retinoblastoma proteins: from sequence and structure analysis to intracellular networks. Front Physiol 4:315CrossRefGoogle Scholar
  10. Himmelbach A, Zierold U, Hensel G et al (2007) A set of modular binary vectors for transformation of cereals. Plant Phys 145:1192–1200. CrossRefGoogle Scholar
  11. Hruz T, Laule O, Szabo G et al (2008) Genevestigator V3: a reference expression database for the meta-analysis of transcriptomes. Adv Bioinform. CrossRefGoogle Scholar
  12. Janski N, Masoud K, Batzenschlager M et al (2012) The GCP3-interacting proteins GIP1 and GIP2 are required for γ-tubulin complex protein localization, spindle integrity, and chromosomal stability. Plant Cell 24:1171–1187. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Juraniec M, Heyman J, Schubert V et al (2016) Arabidopsis COPPER MODIFIED RESISTANCE1/PATRONUS1 is essential for growth adaptation to stress and required for mitotic onset control. New Phytol 209:177–191CrossRefGoogle Scholar
  14. Karimi M, Inzé D. Depicker A (2002) GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7:193–195CrossRefGoogle Scholar
  15. Mahjoubi H, Ebel C, Hanin M (2015) Molecular and functional characterization of the durum wheat TdRL1, a member of the conserved Poaceae RSS1-like family that exhibits features of intrinsically disordered proteins and confers stress tolerance in yeast. Funct Integr Genom 15:717–728. CrossRefGoogle Scholar
  16. Ogawa D, Abe K, Miyao A et al (2011) RSS1 regulates the cell cycle and maintains meristematic activity under stress conditions in rice. Nat Commun 2:278. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Ogawa D, Morita H, Hattori T, Takeda S (2012) Molecular characterization of the rice protein RSS1 required for meristematic activity under stressful conditions. Plant Physiol Biochem 61:54–60CrossRefGoogle Scholar
  18. Pazos F, Pietrosemoli N, García-Martín JA, Solano R (2013) Protein intrinsic disorder in plants. Front Plant Sci 4:363CrossRefGoogle Scholar
  19. Pietrosemoli N, García-Martín JA, Solano R, Pazos F (2013) Genome-wide analysis of protein disorder in Arabidopsis thaliana: implications for plant environmental adaptation. PLoS One 8(2):e55524. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Popov N, Schmitt M, Schulzeck S, Matthies H (1975) Reliable micromethod for determination of the protein content in tissue homogenates. Acta Biol Med Ger 34:1441–1446PubMedGoogle Scholar
  21. Sánchez-Calderón L, Ibarra-Cortés ME, Zepeda-Jazo I (2013) Root development and abiotic stress adaptation. In: Kourosh V (ed) Abiotic stress—plant responses and applications in agriculture. InTech, London. CrossRefGoogle Scholar
  22. Schneider CA, Rasband WS, Eliceiri KW (2012) NIH image to ImageJ: 25 years of Image analysis. Nat Methods 9:671–675CrossRefGoogle Scholar
  23. Shi J, Fu XZ, Peng T et al (2010) Spermine pretreatment confers dehydration tolerance of citrus in vitro plants via modulation of antioxidative capacity and stomatal response. Tree Physiol 30:914–922CrossRefGoogle Scholar
  24. Sun X, Rikkerink EHA, Jones WT, Uversky VN (2013) Multifarious roles of intrinsic disorder in proteins illustrate its broad impact on plant biology. Plant Cell 25:38–55. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Terada R, Asao H, Iida S (2004) A large-scale Agrobacterium-mediated transformation procedure with a strong positive-negative selection for gene targeting in rice (Oryza sativa L.). Plant Cell Rep 22:653–659CrossRefGoogle Scholar
  26. Tian T, Liu Y, Yan H et al (2017) agriGO v2.0: a GO analysis toolkit for the agricultural community, 2017 update. Nucleic Acids Res 45:122–129. CrossRefGoogle Scholar
  27. Velikova V, Yordanov I, Edreva A (2000) Oxidative stress and some antioxidant system in acid rain treated bean plants, protective role of exogenous polyamines. Plant Sci 151:59–66CrossRefGoogle Scholar
  28. Zamariola L, De Storme N, Vannerum K et al (2014) SHUGOSHINs and PATRONUS protect meiotic centromere cohesion in Arabidopsis thaliana. Plant J 77:782–794CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratoire de Biotechnologie et d’Amélioration des PlantesCentre de Biotechnologie de SfaxSfaxTunisia
  2. 2.Institut de biologie moléculaire des plantes, UPR 2357 du CNRSUniversité de StrasbourgStrasbourg CedexFrance
  3. 3.Bioscience and Biotechnology CenterNagoya UniversityNagoyaJapan
  4. 4.Institut Jean-Pierre Bourgin, INRA, AgroParisTech, CNRSUniversité Paris-SaclayVersaillesFrance
  5. 5.Plant Physiology and Functional Genomics Research Unit, Institute of BiotechnologyUniversity of SfaxSfaxTunisia

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