Agrobacterium tumefaciens- Mediated Transformation of Pucciniomycotina Red Yeasts

  • Giuseppe Ianiri
  • Alexander IdnurmEmail author
Part of the Fungal Biology book series (FUNGBIO)


Red yeasts belong to the subphylum Pucciniomycotina, one of the three major lineages in the Basidiomycete fungi. These yeasts, commonly found as saprophytes in the environment, have unique features that make them important for academia, industry, agriculture, and occasionally human health. To understand the genetic basis behind biological processes of interest, the availability of molecular tools for gene mutation plays a crucial role. Here we outline how to perform recently developed protocols of Agrobacterium tumefaciens-mediated transformation to Pucciniomycotina red yeasts. This method can be applied to generate libraries of T-DNA insertional mutants that can be screened for phenotypes of interest, or for targeted gene replacement.


Pucciniomycotina Sporobolomyces sp. Agrobacterium tumefaciens-mediated transformation (AMT) Auxotrophic marker Insertional mutagenesis 



Our research on the Pucciniomycotina fungi has been supported by the University of Missouri Research Board, the US National Science Foundation, and the National Institutes of Health.


  1. Abbott EP, Ianiri G, Castoria R, Idnurm A (2013) Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts. Appl Microbiol Biotechnol 97:283–95PubMedCrossRefGoogle Scholar
  2. Aime MC, Matheny PB, Henk DA, Frieders EM, Nilsson RH, Piepenbring M, McLaughlin DJ, Szabo LJ, Begerow D, Sampaio JP, Bauer R, Weiss M, Oberwinkler F, Hibbett D (2006) An overview of the higher level classification of Pucciniomycotina based on combined analyses of nuclear large and small subunit rDNA sequences. Mycologia 98:896–905PubMedCrossRefGoogle Scholar
  3. Bundock P, den Dulk-Ras A, Beijersbergen A, Hooykaas PJJ (1995) Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J 14:3206–14PubMedPubMedCentralGoogle Scholar
  4. Frandsen RJN (2011) A guide to binary vectors and strategies for targeted genome modification in fungi using Agrobacterium tumefaciens-mediated transformation. J Microbiol Methods 87:247–62PubMedCrossRefGoogle Scholar
  5. Ianiri G, Wright SAI, Castoria R, Idnurm A (2011) Development of resources for the analysis of gene function in Pucciniomycotina red yeasts. Fungal Genet Biol 48:685–95PubMedCrossRefGoogle Scholar
  6. Liu Y, Koh CMJ, Sun L, Hlaing MM, Du M, Peng N, Ji L (2013) Characterization of glyceraldehyde-3-phosphate dehydrogenase gene RtGPD1 and development of genetic transformation method by dominant selection in oleaginous yeast Rhodos-poridium toruloides. Appl Microbiol Biotechnol 97:719–29PubMedCrossRefGoogle Scholar
  7. Michielse CB, Hooykaas PJJ, van den Hondel CAMJJ, Ram AFJ (2005) Agrobacterium-mediated transformation as a tool for functional genomics in fungi. Curr Genet 48:1–17PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  1. 1.School of Biological SciencesUniversity of Missouri-Kansas CityKansas CityUSA

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