MPPI and CFPRG host Cercospora transformation workshop 2014-09-18
Currently there are many fungal genome sequencing projects at FABI. The next step is functional genomics in which hypotheses about gene functions can be tested by making gene knockouts. Prof Dave Berger (MPPI) arranged a lab workshop on protoplast transformation and split-marker gene knockout protocols for Cercospora in the Plant Sciences Complex on his return from Sabbatical with Dr. Burt Bluhm at the Department of Plant Pathology at the University of Arkansas, USA. The workshop was based on protocols published by the Bluhm lab (Ridenour et al. 2012). Transformation of Cercospora with green fluorescent protein (GFP) was used for illustration.
The workshop was attended by a dozen participants from different research groups in the Departments of Plant Science, Genetics, Microbiology and Plant Pathology, as well as a collaborator from the Department of Molecular and Cell Biology at the University of Cape Town. Kara Troglin, a student from the Bluhm lab at the University of Arkansas, who has a wealth of experience with this challenging fungal species, also participated in the workshop. It was held during her four week visit to the group where she worked with postgraduate students on Cercospora culturing techniques, microscopy and PCR analysis. The workshop was successful in generating Cercospora transformants that were hygromycin resistant and GFP positive. Several participants from the MPPI, CFPRG and other groups are applying what they learned in their research projects. A technical “breakthrough” during the workshop was the demonstration by Dr Irene Barnes that protoplasts can be photographed through a microscope eyepiece with an iPhone!
Ridenour JB, Hirsch RL, Bluhm BH (2012) Identifying Genes in Fusarium verticillioides Through Forward and Reverse Genetics. In: Bolton,Thomma (eds) Plant Fungal Pathogens, Humana Press, pp. 457-479.