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Tools for microalgal biotechnology: development of an optimized transformation method for an industrially promising microalga—Tetraselmis chuii

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Abstract

Tetraselmis chuii is a biotechnologically promising green microalga that is commonly used in aquaculture. Development of reliable and easy genetic transformation procedures is essential for the improvement of this microalga. Here, we report the development of a suitable method for the stable genetic transformation of T. chuii via Agrobacterium tumefaciens. Following a statistical experimental design, seven factors were tested for their effect on the transformation efficiency. The effects of temperature, pH, and the interaction between acetosyringone concentration and co-culturing period were statistically significant. These main factors were subsequently optimized to the following values: 27 °C co-culturing temperature, pH 5.0, and 150 μM acetosyringone concentration. Transformation was verified by PCR on different target genes present in the T-DNA as well as by the determination of GUS activity from the reporter gene. Genetic stability of the transformed clones was examined by culturing under non-selective conditions in phleomycin-free medium for 6 months. Transformed clones were stable as indicated by the maintenance of the resistance to phleomycin (PhlR). In summary, an easy and reliable genetic transformation method for T. chuii was developed. The use of A. tumefaciens provides a useful method for the genetic engineering of this biotechnologically promising microalga.

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Acknowledgements

This work was supported by the EnerGeticAl project funded by the Ministerio de Economía y Competitividad (MINECO, project IPT-2011-0842-920000) in collaboration with ENDESA, TECNALIA, AITEMIN, and Universidad de Almería. Yasmeen Dautor was supported by a pre-doctoral fellowship from the Government of Syria.

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Authors and Affiliations

  1. Grupo de investigación “Biotecnología de Productos Naturales” (BIO-279), Centro de Investigación en Biotecnología Agroalimentaria (BITAL), Campus Internacional de Excelencia Agroalimentario (CeiA3), Universidad de Almería, Almería, 04120, Spain

    Patricia Úbeda-Mínguez, Tarik Chileh, Yasmeen Dautor, Federico García-Maroto & Diego López Alonso

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  1. Patricia Úbeda-Mínguez
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  2. Tarik Chileh
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  3. Yasmeen Dautor
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  4. Federico García-Maroto
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  5. Diego López Alonso
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Correspondence to Diego López Alonso.

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Úbeda-Mínguez, P., Chileh, T., Dautor, Y. et al. Tools for microalgal biotechnology: development of an optimized transformation method for an industrially promising microalga—Tetraselmis chuii . J Appl Phycol 27, 223–232 (2015). https://doi.org/10.1007/s10811-014-0306-6

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  • DOI: https://doi.org/10.1007/s10811-014-0306-6

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