Skip to main content
Log in

Identification of differentially expressed genes in Alternanthera philoxeroides under drought stress using suppression subtractive hybridization

  • Research Papers
  • Published:
Russian Journal of Plant Physiology Aims and scope Submit manuscript

Abstract

The current study was conducted by using suppression subtractive hybridization (SSH) to identify the differentially expressed genes in Alternanthera philoxeroides under drought stress and thus to explore the molecular mechanisms of drought tolerance. The mRNA was extracted from the roots of drought-treated and well-watered A. philoxeroides to construct SSH cDNA library. Positive clones were selected for sequencing and further analyzed by BLAST for screening non-redundant and homologous expressed sequence tags (ESTs). Then these ESTs were put into the Gene Ontology database for functional annotation and Kyoto Encylopedia of Genes and Genomes (KEGG) for metabolic pathways analysis. Four cDNA fragments, ZFP (zinc finger protein), HSP70 (heat shock protein 70), CAT (catalase), and TPS (trehalose-6-phosphate synthase), were randomly chosen for RT-PCR analysis. In the SSH cDNA library, 286 positive clones picked up randomly were sequenced and finally 269 sequences were available. After cluster analysis of the ESTs, 82 unigenes were obtained, in which 63 genes displayed a high homology to known sequences. KEGG analysis found that these genes were involved in 14 metabolic pathways, such as the pathways of plant hormone signal transduction and biosynthesis of secondary metabolites. The expressions of the above four cDNA fragments were all up-regulated in A. philoxeroides under drought stress. This study presented a basis forbstudying the drought tolerance mechanism of A. philoxeroides, which provided a theoretical basis for managing the spread of this plant.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

CAT:

catalase

EST:

expressed sequence tag

HSP70:

heat shock protein 70

KEGG:

Kyoto Encylopedia of Genes and Genomes

SSH:

suppression subtractive hybridization

TPS:

trehalose-6-phosphate synthase

ZFP:

zinc finger protein

References

  1. Buckingham, G.R., Biological control of alligatorweed, Alternanthera philoxeroides, the world’s first aquatic weed success story, Castanea, 1996, vol. 61, pp. 232–243.

    Google Scholar 

  2. Julien, M.H., Skarratt, B., and Maywald, G., Potential geographical distribution of alligator weed and its biological control by Agasicles hygrophila, J. Aquat. Plant Manage., 1995, vol. 33, pp. 55–60.

    Google Scholar 

  3. Gao, J., Xiao, Q., Yin, L., and He, G., Isolation of cDNA clones for genes up-regulated in droughttreated Alternanthera philoxeroides root, Mol. Biol. Rep., 2008, vol. 35, pp. 485–488.

    Article  CAS  PubMed  Google Scholar 

  4. Shinozaki, K. and Yamaguchi-Shinozaki, K., Gene networks involved in drought stress response and tolerance, J. Exp. Bot., 2007, vol. 58, pp. 221–227.

    Article  CAS  PubMed  Google Scholar 

  5. Yamamoto, K. and Sasaki, T., Large-scale EST sequencing in rice, Plant Mol. Biol., 1997, vol. 35, pp. 135–144.

    Article  CAS  PubMed  Google Scholar 

  6. Li, J. and Ye, W.H., Genetic diversity of alligator weed ecotypes is not the reason for their different responses to biological control, Aquat. Bot., 2006, vol. 85, pp. 155–158.

    Article  Google Scholar 

  7. Wong, Y.Y., Ho, C.L., Nguyen, P.D., Teo, S.S., Harikrishna, J.A., Rahim, R.A., and Wong, M.C., Isolation of salinity tolerant genes from the mangrove plant, Bruguiera cylindrical by using suppression subtractive hybridization (SSH) and bacterial functional screening, Aquat. Bot., 2007, vol. 86, pp. 117–122.

    Article  CAS  Google Scholar 

  8. Gronover, C.S., Schorn, C., and Tudzynski, B., Identification of Botrytis cinerea genes up-regulated during infection and controlled by the Gα subunit BCG1 using suppression subtractive hybridization (SSH), Mol. Plant-Microbe Interact., 2004, vol. 17, pp. 537–546.

    Article  CAS  Google Scholar 

  9. Wu, Y., Wang, Q., Ma, Y., and Chu, C., Isolation and expression analysis of salt up-regulated ESTs in upland rice using PCR-based subtractive suppression hybridization method, Plant Sci., 2005, vol. 168, pp. 847–853.

    Article  CAS  Google Scholar 

  10. Munnik, T. and Meijer, H.J., Osmotic stress activates distinct lipid and MAPK signalling pathways in plants, FEBS Lett., 2001, vol. 498, pp. 172–178.

    Article  CAS  PubMed  Google Scholar 

  11. Xu, H., Li, K., Yang, F., Shi, Q., and Wang, X., Overexpression of CsNMAPK in tobacco enhanced seed germination under salt and osmotic stresses, Mol. Biol. Rep., 2010, vol. 37, pp. 3157–3163.

    Article  CAS  PubMed  Google Scholar 

  12. Xiong, L., Schumaker, K.S., and Zhu, J.K., Cell signaling during cold, drought, and salt stress, Plant Cell Online, 2002, vol. 14, pp. S165–S183.

    Article  CAS  Google Scholar 

  13. Xu, G.Y., Rocha, P.S., Wang, M.L., Xu, M.L., Cui, Y.C., Li, L.Y., Zhu, Y.X., and Xia, X., A novel rice calmodulinlike gene, OsMSR2, enhances drought and salt tolerance and increases ABA sensitivity in Arabidopsis, Planta, 2011, vol. 234, pp. 47–59.

    Article  CAS  PubMed  Google Scholar 

  14. Takatsuji, H., Zinc-finger transcription factors in plants, Cell Mol. Life Sci., 1998, vol. 54, pp. 582–596.

    Article  CAS  PubMed  Google Scholar 

  15. Sakamoto, H., Maruyama, K., Sakuma, Y., Meshi, T., Iwabuchi, M., Shinozaki, K., and Yamaguchi-Shinozaki, K., Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and high-salinity stress conditions, Plant Physiol., 2004, vol. 136, pp. 2734–2746.

    CAS  Google Scholar 

  16. Ritossa, F., A new puffing pattern induced by temperature shock and DNP in Drosophila, Experientia, 1962, vol. 18, pp. 571–573.

    Article  CAS  Google Scholar 

  17. Wang, W., Vinocur, B., Shoseyov, O., and Altman, A., Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response, Trends Plant Sci., 2004, vol. 9, pp. 244–252.

    Article  CAS  PubMed  Google Scholar 

  18. Liu, C.C., Li, C.M., Liu, B.G., Ge, S.J., Dong, X.M., Li, W., Zhu, H.Y., Wang, B.C., and Yang, C.P., Genome-wide identification and characterization of a dehydrin gene family in poplar (Populus trichocarpa), Plant Mol. Biol. Rep., 2012, vol. 30, pp. 848–859.

    Article  CAS  Google Scholar 

  19. Asada, K. and Takahashi, M., Production and scavenging of active oxygen in photosynthesis, Photoinhibition Topics in Photosynthesis. V. 9, Kyle, D.J., Osmond, C.B., and Arntzen, C.J, Eds., Amsterdam: Elsevier, 1987, pp. 227–287.

    Google Scholar 

  20. Bowler, C., Montagu, M.V., and Inze, D., Superoxide dismutase and stress tolerance, Annu. Rev. Plant Biol., 1992, vol. 43, pp. 83–116.

    Article  CAS  Google Scholar 

  21. Willekens, H., Chamnongpol, S., Davey, M., Schraudner, M., Langebartels, C., van Montagu, M., Inzé, D., and van Camp, W., Catalase is a sink for H2O2 and is indispensable for stress defense in C3 plants, EMBO J., 1997, vol. 16, pp. 4806–4816.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Mittler, R., Oxidative stress, antioxidants and stress tolerance, Trends Plant Sci., 2002, vol. 7, pp. 405–410.

    Article  CAS  PubMed  Google Scholar 

  23. Garg, A.K., Kim, J.K., Owens, T.G., Ranwala, A.P., Do Choi, Y., Kochian, L.V., and Wu, R.J., Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses, Proc. Natl. Acad. Sci. USA, 2002, vol. 99, pp. 15 898–15 903.

    Article  CAS  Google Scholar 

  24. Romero, C., Bellés, J.M., Vayá, J.L., Serrano, R., and Culiáñez-Macià, F.A., Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance, Planta, 1997, vol. 201, pp. 293–297.

    Article  CAS  PubMed  Google Scholar 

  25. Kosmas, S.A., Argyrokastritis, A., Loukas, M.G., Eliopoulos, E., Tsakas, S., and Kaltsikes, P.J., Isolation and characterization of drought-related trehalose 6-phosphate-synthase gene from cultivated cotton (Gossypium hirsutum L.), Planta, 2006, vol. 223, pp. 329–339.

    Article  CAS  PubMed  Google Scholar 

  26. Bray, E.A., Plant responses to water deficit, Trends Plant Sci., 1997, vol. 2, pp. 48–54.

    Article  Google Scholar 

  27. Salekdeh, G., Siopongco, J., Wade, L.J., Ghareyazie, B., and Bennett, J., Proteomic analysis of rice leaves during drought stress and recovery, Proteomics, 2002, vol. 2, pp. 1131–1145.

    Article  CAS  PubMed  Google Scholar 

  28. Jorge, I., Navarro, R.M., Lenz, C., Ariza, D., and Jorrín, J., Variation in the holm oak leaf proteome at different plant developmental stages, between provenances and in response to drought stress, Proteomics, 2006, vol. 6, pp. S207–S214.

    Article  PubMed  Google Scholar 

  29. Plomion, C., Lalanne, C., Claverol, S., Meddour, H., Kohler, A., Bogeat-Triboulot, M.B., Barre, A., Le Provost, G., Dumazet, H., and Jacob, D., Mapping the proteome of poplar and application to the discovery of drought-stress responsive proteins, Proteomics, 2006, vol. 6, pp. 6509–6527.

    Article  CAS  PubMed  Google Scholar 

  30. Bogeat-Triboulot, M.B., Brosché, M., Renaut, J., Jouve, L., Le Thiec, D., Fayyaz, P., Vinocur, B., Witters, E., Laukens, K., and Teichmann, T., Gradual soil water depletion results in reversible changes of gene expression, protein profiles, ecophysiology, and growth performance in Populus euphratica, a poplar growing in arid regions, Plant Physiol., 2007, vol. 143, pp. 876–892.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R. Y. Ma.

Additional information

This text was submitted by the authors in English.

Joint first authors, Dong Jia and Bing Zhang, contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jia, D., Zhang, B., Zhang, P.P. et al. Identification of differentially expressed genes in Alternanthera philoxeroides under drought stress using suppression subtractive hybridization. Russ J Plant Physiol 62, 93–100 (2015). https://doi.org/10.1134/S1021443715010094

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1021443715010094

Keywords

Navigation