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Identification of Copper-Induced Genes in the Marine Alga Ulva compressa (Chlorophyta)

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Abstract

In order to identify genes/proteins involved in copper tolerance, the marine alga Ulva compressa was cultivated with 10 μM copper for 3 days. The activities of antioxidant enzymes ascorbate peroxidase (AP), peroxiredoxin (PRX), thioredoxin (TRX), and glutathione-S-transferase (GST) and the level of lipoperoxides were determined in the alga cultivated with and without copper addition. Antioxidant enzyme activities and lipoperoxides level increased in response to copper excess, indicating that the alga was under oxidative stress. A cDNA library was prepared using U. compressa cultivated with 10 μM copper for 3 days. A total of 3 × 104 clones were isolated and 480 clones were sequenced, resulting in 235 non-redundant ESTs, of which 104 encode proteins with known functions. Among them, we identified proteins involved in (1) antioxidant metabolism such as AP, PRX, TRX, GST, and metalothionein (MET), (2) signal transduction, such as calmodulin (CAM), (3) calcium-dependent protein kinase (CDPK) and nucleoside diphosphate kinase (NDK), (4) gene expression, (5) protein synthesis and degradation, and (6) chloroplast and mitochondria electron transport chains. Half of the identified proteins are potentially localized in organelles. The relative level of 18 genes, including those coding for AP, PRX, TRX, GST, MET, CAM, CDPK, and NDK were determined by quantitative RT-PCR in the alga cultivated with 10 μM copper for 0 to 7 days. Transcript levels increased in response to copper stress and most of them reached a maximum at days 3 and 5. Thus, the selected genes are induced by copper stress and they are probably involved in copper acclimation and tolerance.

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Acknowledgments

This work was supported by the research programs Marine Genomics Chile from CONICYT to JAC and AM, DICYT-USACH and FONDECYT 1085041 to AM, FONDAP 1501-0001 subprogram 7 from CONICYT and ICA to JAC and FONDECYT 11085019 to LC. We thank Marine Genomics Europe for the fellowship to LC, Dr. Richard Reinhardt and Dr. Michael Kube from the Max-Planck institute for Molecular Genetics (Berlin, Germany) where the cDNA library was prepared and the ESTs analysis was performed.

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Authors and Affiliations

  1. Departamento de Ecología, Center for Advanced Studies in Ecology and Biodiversity, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, 6513677, Santiago, Chile

    Loretto Contreras-Porcia & Juan A. Correa

  2. Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile, Casilla 40 correo 33, Santiago, Chile

    Geraldine Dennett, Alberto González, Eva Vergara, Cristóbal Medina & Alejandra Moenne

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  1. Loretto Contreras-Porcia
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  2. Geraldine Dennett
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  3. Alberto González
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  4. Eva Vergara
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Correspondence to Alejandra Moenne.

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Table 1

PCR primers used for quantitative RT-PCR amplification of U. compressa gene transcripts (DOC 64 kb)

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Contreras-Porcia, L., Dennett, G., González, A. et al. Identification of Copper-Induced Genes in the Marine Alga Ulva compressa (Chlorophyta). Mar Biotechnol 13, 544–556 (2011). https://doi.org/10.1007/s10126-010-9325-8

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