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Isolation, molecular characterization and functional analysis of OeMT2, an olive metallothionein with a bioremediation potential

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Abstract

Metallothioneins are essential in plants for metal detoxification in addition to their other roles in plant life cycle. This study reports the characterization of an olive (Olea europaea L. cv. Ayvalik) metallothionein with respect to molecular and functional properties. A cDNA encoding a type 2 metallothionein from olive was isolated from a leaf cDNA library, characterized and named OeMT2 after its molecular and functional properties. OeMT2 was expressed in Escherichia coli, and a single protein band was confirmed by protein gel blot analysis. Metal tolerance ability of bacterial cells expressing OeMT2 was determined against 0.2 mM CdCl2, 0.4 mM CdCl2 and 1 mM CuSO4 in the growth medium. Metal ion contents of bacterial cells expressing OeMT2 were measured by ICP. Metal tolerance assays and ICP measurements suggested that OeMT2 effectively binds Cu and Cd. Molecular analysis of OeMT2 revealed two introns, three exons, a short 3′ UTR and a long 5′ UTR. Comparing the genomic sequences from 14 olive cultivars revealed OeMT2 had both intron and exon polymorphisms dividing the cultivars into three groups. Real-time PCR analysis demonstrated that OeMT2 expresses more or less the same amounts in all tissues of the olive tree examined. The genomic copy number of OeMT2 was also determined employing real-time PCR which suggested a single copy gene in the olive genome while three other MT2 members were determined from the draft olive genome sequences of Ayvalik cultivar and that of wild olive. This is the first report on molecular and functional characterization of an olive metallothionein and shows that OeMT2 expressed in E. coli has the capability of effectively binding toxic heavy metals. This may suggest that OeMT2 plays an important role in metal homeostasis in addition to a good potential for environmental and industrial usage.

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Acknowledgments

We gratefully acknowledge TUBITAK for supporting this work with the grant numbered 110O108. We also thank BUTAM for providing ICP analysis with us.

Conflict of interest

Authors declare no conflict of interest.

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Authors and Affiliations

  1. Biyoloji Bölümü (Department of Biology), Fen Edebiyat Fakültesi (College of Arts and Sciences), Balikesir University, Cagis, 10145, Balikesir, Turkey

    Ekrem Dundar & Görkem Deniz Sonmez

  2. Biyoloji Bölümü (Department of Biology), Fen Fakültesi (Faculty of Science), Cankiri Karatekin University, Ballica, 18100, Cankiri, Turkey

    Turgay Unver

Authors
  1. Ekrem Dundar
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  2. Görkem Deniz Sonmez
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  3. Turgay Unver
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Correspondence to Ekrem Dundar.

Additional information

Communicated by S. Hohmann.

Data Archiving Statements

The OeMT2 sequence has been submitted to NCBI-GenBank with the accession number GW574261. The accession numbers of the other OeMT2 members will be provided once the olive genome is constructed and deposited to the GenBank.

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Dundar, E., Sonmez, G.D. & Unver, T. Isolation, molecular characterization and functional analysis of OeMT2, an olive metallothionein with a bioremediation potential. Mol Genet Genomics 290, 187–199 (2015). https://doi.org/10.1007/s00438-014-0908-3

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  • DOI: https://doi.org/10.1007/s00438-014-0908-3

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