Mycobiology

The Korean Society of Mycology (한국균학회)
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Agrobacterium-Mediated Co-transformation of Multiple Genes in Metarhizium robertsii

  • Padilla-Guerrero, Israel Enrique (Division of Natural and Exact Sciences, Department of Biology, University of Guanajuato, Campus Guanajuato) ;
  • Bidochka, Michael J. (Department of Biological Sciences, Brock University)
  • Received : 2017.05.10
  • Accepted : 2017.06.11
  • Published : 2017.06.30
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Abstract

Fungi of the Metarhizium genus are a very versatile model for understanding pathogenicity in insects and their symbiotic relationship with plants. To establish a co-transformation system for the transformation of multiple M. robertsii genes using Agrobacterium tumefaciens, we evaluated whether the antibiotic nourseothricin has the same marker selection efficiency as phosphinothricin using separate vectors. Subsequently, in the two vectors containing the nourseothricin and phosphinothricin resistance cassettes were inserted eGFP and mCherry expression cassettes, respectively. These new vectors were then introduced independently into A. tumefaciens and used to transform M. robertsii either in independent events or in one single co-transformation event using an equimolar mixture of A. tumefaciens cultures. The number of transformants obtained by co-transformation was similar to that obtained by the individual transformation events. This method provides an additional strategy for the simultaneous insertion of multiple genes into M. robertsii.

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References

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