Skip to main content
Log in

Functional analysis of Triticum durum type 1 metallothionein gene (dMT) in response to varying levels of cadmium

  • Original Article
  • Published:
Indian Journal of Plant Physiology Aims and scope Submit manuscript

Abstract

The effect of varying levels of cadmium and correlated changes on the expression level of a type 1 metallothionein gene (dMT) were investigated in Triticum durum cv. Balçalı-85. Increasing the cadmium concentration resulted in a decrease in the dry weights of roots and shoots, and the effect was stronger in roots. Roots also showed a higher capacity to accumulate cadmium. Southern blot analyses revealed that the dMT gene, delineated by two exons and a non-coding intron region, exists at a single locus in the T. durum genome. Changes in dMT gene expression during cadmium exposure were monitored by two approaches. Northern blot analyses showed that the transcript level in roots increased upon treatment with increasing cadmium, which was quantified by qRT-PCR as 4.5 fold of the base level at 10 μM Cd. These results show a positive correlation between cadmium exposure and expression of dMT gene in durum wheat, and will provide a basis for studies on the role of type 1 metallothioneins in cadmium response.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Akashi, K., Nishimura, N., Ishida, Y., & Yokota, A. (2004). Potent hydroxyl radical-scavenging activity of drought-induced type-2 metallothionein in wild watermelon. Biochemical and Biophysical Research Communications, 323, 72–78.

    Article  CAS  PubMed  Google Scholar 

  • Bilecen, K., Ozturk, U. H., Duru, A. D., Sutlu, T., Petoukhov, M. V., Svergun, D. I., et al. (2005). Triticum durum metallothionein. Isolation of the gene and structural characterization of the protein using solution scattering and molecular modeling. Journal of Biological Chemistry, 280, 13701–13711.

    Article  CAS  PubMed  Google Scholar 

  • Binz, P. A., & Kagi, J. H. R. (1999). Metallothionein: Molecular evolution and classification. In C. Klaasen (Ed.), Metallothionein IV (pp. 7–13). Basel: Birkhäuser Verlag.

    Chapter  Google Scholar 

  • Brkljacić, J. M., Samardzić, J. T., Timotijević, G. S., & Maksimović, V. R. (2004). Expression analysis of buckwheat (Fagopyrum esculentum Moench) metallothionein-like gene (MT3) under different stress and physiological conditions. Journal of Plant Physiology, 161, 741–746.

    Article  PubMed  Google Scholar 

  • Butt, A., Mousley, C., Morris, K., Beynon, J., Can, C., Holub, E., et al. (1998). Differential expression of a senescence-enhanced metallothionein gene in Arabidopsis in response to isolates of Peronospora parasitica and Pseudomonas syringae. The Plant Journal, 16, 209–221.

    Article  CAS  PubMed  Google Scholar 

  • Cebeci, Ö., Köktürk, B., Ergen, N., Öztür, L., Çakmak, İ., & Budak, H. (2008). Differential expression of wheat transcriptomes in response to varying cadmium concentrations. Biologia Plantarum, 52, 703–708.

    Article  CAS  Google Scholar 

  • Chang, T., Liu, X., Xu, H., Meng, K., Chen, S., & Zhu, Z. (2004). A metallothionein-like gene htMT2 strongly expressed in internodes and nodes of Helianthus tuberosus and effects of metal ion treatment on its expression. Planta, 218, 449–455.

    Article  CAS  PubMed  Google Scholar 

  • Charbonnel-Campaa, L., Lauga, B., & Combes, D. (2000). Isolation of a type 2 metallothionein-like gene preferentially expressed in the tapetum in Zea mays. Gene, 254, 199–208.

    Article  CAS  PubMed  Google Scholar 

  • Chatthai, M., Kaukinen, K. H., Tranbarger, T. J., Gupta, P. K., & Misra, S. (1997). The isolation of a novel metallothionein-related cDNA expressed in somatic and zygotic embryos of Douglas-fir: Regulation by ABA, osmoticum, and metal ions. Plant Molecular Biology, 34, 243–254.

    Article  CAS  PubMed  Google Scholar 

  • Chen, H. J., Hou, W. C., Yang, C. Y., Huang, D. J., Liu, J. S., & Lin, Y. H. (2003). Molecular cloning of two metallothionein-like protein genes with differential expression patterns from sweet potato (Ipomoea batatas) leaves. Journal of Plant Physiology, 160, 547–555.

    Article  CAS  PubMed  Google Scholar 

  • Cho, S. H., Hoang, Q. T., Kim, Y. Y., Shin, H. Y., Ok, S. H., Bae, J. M., et al. (2006). Proteome analysis of gametophores identified a metallothionein involved in various abiotic stress responses in Physcomitrella patens. Plant Cell Reports, 25, 475–488.

    Article  CAS  PubMed  Google Scholar 

  • Choi, D., Kim, H. M., Yun, H. K., Park, J. A., Kim, W. T., & Bok, S. H. (1996). Molecular cloning of a metallothionein-like gene from Nicotiana glutinosa L. and its induction by wounding and tobacco mosaic virus infection. Plant Physiology, 112, 353–359.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Clemens, S. (2006). Toxic metal accumulation, responses to exposure and mechanisms of tolerance in plants. Biochimie, 88, 1707–1719.

    Article  CAS  PubMed  Google Scholar 

  • Cobbett, C., & Goldsbrough, P. (2002). Phytochelatins and metallothioneins: Roles in heavy metal detoxification and homeostasis. Annual Review of Plant Biology, 53, 159–182.

    Article  CAS  PubMed  Google Scholar 

  • Davies, C., & Robinson, S. P. (2000). Differential screening indicates a dramatic change in mRNA profiles during grape berry ripening. Cloning and characterization of cDNAs encoding putative cell wall and stress response proteins. Plant Physiology, 122, 803–812.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • de Framond, A. J. (1991). A metallothionein-like gene from maize (Zea mays). Cloning and characterization. FEBS Letters, 290, 103–106.

    Article  PubMed  Google Scholar 

  • Di Toppi, L. S., & Gabbrielli, R. (1999). Response to cadmium in higher plants. Environmental and Experimental Botany, 41, 105–130.

    Article  Google Scholar 

  • Evans, K. M., Gatehouse, J. A., Lindsay, W. P., Shi, J., Tommey, A. M., & Robinson, N. J. (1992). Expression of the pea metallothionein-like gene PsMTA in Escherichia coli and Arabidopsis thaliana and analysis of trace metal ion accumulation: Implications for PsMTA function. Plant Molecular Biology, 20, 1019–1028.

    Article  CAS  PubMed  Google Scholar 

  • Freisinger, E. (2008). Plant MTs-long neglected members of the metallothionein superfamily. Dalton Transactions, 47, 6663–6675.

    Article  Google Scholar 

  • Grant, C. A., Buckley, W. T., Bailey, L. D., & Selles, F. (1998). Cadmium accumulation in crops. Canadian Journal of Plant Science, 78, 1–17.

    Article  CAS  Google Scholar 

  • Guo, W. J., Bundithya, W., & Goldsbrough, P. B. (2003). Characterization of the Arabidopsis metallothionein gene family: Tissue-specific expression and induction during senescence and in response to copper. New Phytologist, 159, 369–381.

    Article  CAS  Google Scholar 

  • Hsieh, H. M., Liu, W. K., Chang, A., & Huang, P. C. (1996). RNA expression patterns of a type 2 metallothionein-like gene from rice. Plant Molecular Biology, 32, 525–529.

    Article  CAS  PubMed  Google Scholar 

  • Hsieh, H. M., Liu, W. K., & Huang, P. C. (1995). A novel stress-inducible metallothionein-like gene from rice. Plant Molecular Biology, 28, 381–389.

    Article  CAS  PubMed  Google Scholar 

  • Hudspeth, R. L., Hobbs, S. L., Anderson, D. M., Rajasekaran, K., & Grula, J. W. (1996). Characterization and expression of metallothionein-like genes in cotton. Plant Molecular Biology, 31, 701–705.

    Article  CAS  PubMed  Google Scholar 

  • Jin, S., Cheng, Y., Guan, Q., Liu, D., Takano, T., & Liu, S. (2006). A metallothionein-like protein of rice (rgMT) functions in E. coli and its gene expression is induced by abiotic stresses. Biotechnology Letters, 28, 1749–1753.

    Article  CAS  PubMed  Google Scholar 

  • Kim, S. H., Jeong, J. C., Ahn, Y. O., Lee, H. S., & Kwak, S. S. (2014). Differential responses of three sweetpotato metallothionein genes to abiotic stress and heavy metals. Molecular Biology Reports, 41, 6957–6966.

    Article  CAS  PubMed  Google Scholar 

  • Kısa, D., Öztürk, L., & Tekin, S. (2016). Gene expression analysis of metallothionein and mineral elements uptake in tomato (Solanum lycopersicum) exposed to cadmium. Journal of Plant Research, 129, 989–995.

    Article  PubMed  Google Scholar 

  • Klaassen, C. D., Liu, J., & Choudhuri, S. (1999). Metallothionein: An intracellular protein to protect against cadmium toxicity. Annual Review of Pharmacology and Toxicology, 39, 267–294.

    Article  CAS  PubMed  Google Scholar 

  • Kokturk, B. (2006). Cadmium uptake and antioxidative enzyme in durum wheat cultivars in response to increasing Cd application. Resource document. http://agronomysocietyofpakistan.yolasite.com/resources/thesis%20cadmium%20full.pdf.

  • Lee, J., Shim, D., Song, W. Y., Hwang, I., & Lee, Y. (2004). Arabidopsis metallothioneins 2a and 3 enhance resistance to cadmium when expressed in Vicia faba guard cells. Plant Molecular Biology, 54, 805–815.

    Article  CAS  PubMed  Google Scholar 

  • Leszczyszyn, O. I., Imam, H. T., & Blindauer, C. A. (2013). Diversity and distribution of plant metallothioneins: A review of structure, properties and functions. Metallomics, 5, 1146–1169.

    Article  CAS  PubMed  Google Scholar 

  • Muller, P. Y., Janovjak, H., Miserez, A. R., & Dobbie, Z. (2002). Processing of gene expression data generated by quantitative real-time RT-PCR. Biotechniques, 32, 1372–1374, 1376, 1378–1379.

  • Murray, M. G., & Thompson, W. F. (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research, 8, 4321–4325.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Navabpour, S., Morris, K., Allen, R., Harrison, E., A-H-Mackerness, S., & Buchanan-Wollaston, V. (2003). Expression of senescence-enhanced genes in response to oxidative stress. Journal of Experimental Botany, 54, 2285–2292.

    Article  CAS  PubMed  Google Scholar 

  • Nezhad, R. M., Shahpiri, A., & Mirlohi, A. (2013). Heterologous expression and metal-binding characterization of a type 1 metallothionein isoform (OsMTI-1b) from rice (Oryza sativa). Protein Journal, 32, 131–137.

    Article  CAS  PubMed  Google Scholar 

  • Nishiuchi, S., Liu, S., & Takano, T. (2007). Isolation and characterization of a metallothionein-1 protein in Chloris virgata Swartz that enhances stress tolerances to oxidative, salinity and carbonate stress in Saccharomyces cerevisiae. Biotechnology Letters, 29, 1301–1305.

    Article  CAS  PubMed  Google Scholar 

  • Ozturk, L., Eker, S., Ozkutlu, F., & Cakmak, I. (2003). Effect of cadmium on growth and concentration of cadmium, ascorbic acid and sulphydryl groups in durum wheat cultivars. Turkish Journal of Agriculture and Forestry, 27, 161–168.

    CAS  Google Scholar 

  • Palmiter, R. D. (1998). The elusive function of metallothioneins. Proceedings of the National Academy of Sciences of the United States of America, 95, 8428–8430.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Paradiso, A., Berardino, R., de Pinto, M. C., Sanità di Toppi, L., Storelli, M. M., Tommasi, F., et al. (2008). Increase in ascorbate-glutathione metabolism as local and precocious systemic responses induced by cadmium in durum wheat plants. Plant and Cell Physiology, 49, 362–374.

    Article  CAS  PubMed  Google Scholar 

  • Polle, A., & Schützendübel, S. (2003). Heavy metal signalling in plants: Linking cellular and organismic responses. In H. Hirt & K. Shinozaki (Eds.), Plant responses to abiotic stress (Vol. 4, pp. 187–215). Berlin: Springer.

    Chapter  Google Scholar 

  • Quan, X. Q., Wang, Z. L., Zhang, H., & Bi, Y. P. (2008). Cloning and characterization of TsMT3, a type 3 metallothionein gene from salt cress (Thellungiella salsuginea). DNA Sequence, 19, 340–346.

    Article  CAS  PubMed  Google Scholar 

  • Rai, V., Khatoon, S., Bisht, S. S., & Mehrotra, S. (2005). Effect of cadmium on growth, ultramorphology of leaf and secondary metabolites of Phyllanthus amarus Schum. and Thonn. Chemosphere, 61, 1644–1650.

    Article  CAS  PubMed  Google Scholar 

  • Rauser, W. E. (1999). Structure and function of metal chelators produced by plants: The case for organic acids, amino acids, phytin, and metallothioneins. Cell Biochemistry and Biophysics, 31, 19–48.

    Article  CAS  PubMed  Google Scholar 

  • Reynolds, T. L., & Crawford, R. L. (1996). Changes in abundance of an abscisic acid-responsive, early cysteine-labeled metallothionein transcript during pollen embryogenesis in bread wheat (Triticum aestivum). Plant Molecular Biology, 32, 823–829.

    Article  CAS  PubMed  Google Scholar 

  • Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Snowden, K. C., Richards, K. D., & Gardner, R. C. (1995). Aluminum-induced genes (induction by toxic metals, low calcium, and wounding and pattern of expression in root tips). Plant Physiology, 107, 341–348.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vašák, M., & Kägi, J. H. R. (1994). Metallothioneins. In R. B. King (Ed.), Encyclopedia of inorganic chemistry (pp. 2229–2241). New York: Wiley.

    Google Scholar 

  • Wong, H. L., Sakamoto, T., Kawasaki, T., Umemura, K., & Shimamoto, K. (2004). Down-regulation of metallothionein, a reactive oxygen scavenger, by the small GTPase OsRac1 in rice. Plant Physiology, 135, 1447–1456.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xue, T., Li, X., Zhu, W., Wu, C., Yang, G., & Zheng, C. (2009). Cotton metallothionein GhMT3a, a reactive oxygen species scavenger, increased tolerance against abiotic stress in transgenic tobacco and yeast. Journal of Experimental Botany, 60, 339–349.

    Article  CAS  PubMed  Google Scholar 

  • Yang, Z., Wu, Y., Li, Y., Ling, H. Q., & Chu, C. (2009). OsMT1a, a type 1 metallothionein, plays the pivotal role in zinc homeostasis and drought tolerance in rice. Plant Molecular Biology, 70, 219–229.

    Article  CAS  PubMed  Google Scholar 

  • Yang, M., Zhang, F., Wang, F., Dong, Z., Cao, Q., & Chen, M. (2015). Characterization of a type 1 metallothionein gene from the stresses-tolerant plant Ziziphus jujuba. International Journal of Molecular Sciences, 16(8), 16750–16762.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yourtchi, M. S., & Bayat, H. R. (2013). Effect of cadmium toxicity on growth, cadmium accumulation and macronutrient content of durum wheat (Dena CV.). International Journal of Agriculture and Crop Sciences, 6, 1099–1103.

    CAS  Google Scholar 

  • Yu, L. H., Umeda, M., Liu, J. Y., Zhao, N. M., & Uchimiya, H. (1998). A novel MT gene of rice plants is strongly expressed in the node portion of the stem. Gene, 206, 29–35.

    Article  CAS  PubMed  Google Scholar 

  • Yuan, J., Chen, D., Ren, Y., Zhang, X., & Zhao, J. (2008). Characteristic and expression analysis of a metallothionein gene, OsMT2b, down-regulated by cytokinin suggests functions in root development and seed embryo germination of rice. Plant Physiology, 146, 1637–1650.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou, J., & Goldsbrough, P. B. (1994). Functional homologs of fungal metallothionein genes from Arabidopsis. Plant Cell, 6, 875–884.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou, J., & Goldsbrough, P. B. (1995). Structure, organization and expression of the metallothionein gene family in Arabidopsis. Molecular and General Genetics, 248, 318–328.

    Article  CAS  PubMed  Google Scholar 

  • Zhou, G. K., Xu, Y. F., & Liu, J. Y. (2005). Characterization of a rice class II metallothionein gene: Tissue expression patterns and induction in response to abiotic factors. Journal of Plant Physiology, 162, 686–696.

    Article  CAS  PubMed  Google Scholar 

  • Zimeri, A. M., Dhankher, O. P., McCaig, B., & Meagher, R. B. (2005). The plant MT1 metallothioneins are stabilized by binding cadmiums and are required for cadmium tolerance and accumulation. Plant Molecular Biology, 58, 839–855.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr Atilla Yazici, Veli Bayir for help with ICP measurement and Ozay Ozgur Gokmen for plant physiology experiments in Sabanci University (Istanbul/Turkey).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Filiz Yesilirmak.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yesilirmak, F., Ozturk Gökçe, Z.N., Metin, B. et al. Functional analysis of Triticum durum type 1 metallothionein gene (dMT) in response to varying levels of cadmium. Ind J Plant Physiol. 23, 140–147 (2018). https://doi.org/10.1007/s40502-017-0318-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40502-017-0318-8

Keywords

Navigation