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Functional Analysis of Arbuscular Mycorrhizal Fungal Genes in Yeast

  • Elisabeth TamayoEmail author
  • Tamara Gómez-Gallego
  • Nuria Ferrol
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2146)

Abstract

The obligate symbiotic nature of arbuscular mycorrhizal (AM) fungi makes extremely difficult their genetic manipulation or transformation. For this reason, a heterologous system has been traditionally used for functional analysis of AM fungal genes, being the budding yeast Saccharomyces cerevisiae an organism suitable for this purpose. Here we present the yeast methods required for the functional analysis of AM fungal genes, including protocols for yeast transformation, heterologous gene expression, functional complementation assays, preparation of yeast extracts, and subcellular localization of the encoded protein.

Key words

Arbuscular mycorrhizal fungus Functional characterization Heterologous gene expression Iron transport Saccharomyces cerevisiae Subcellular localization Yeast transformation 
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Notes

Acknowledgments

Tamara Gómez-Gallego was supported by a PhD contract (FPI) from the MINECO and Elisabeth Tamayo was supported by a postdoctoral fellowship from the Alfonso Martin Escudero Foundation. We are grateful to Andrew Dancis for the laboratory protocol of iron transport assays. Research was funded by project RTI2018-098756-B-I00 (MCIU/AEI/FEDER, UE). Elisabeth Tamayo and Tamara Gómez-Gallego contributed equally to this work.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Elisabeth Tamayo
    • 1
    Email author
  • Tamara Gómez-Gallego
    • 2
  • Nuria Ferrol
    • 2
  1. 1.Molecular Phytopathology DepartmentBotanical Institute, Karlsruhe Institute of Technology (KIT)KarlsruheGermany
  2. 2.Department of Soil Microbiology and Symbiotic SystemsEstación Experimental del Zaidín, CSICGranadaSpain

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