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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! 

Reference

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.

 

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Human ZR, Crous CJ, Roets F, Venter SN, Wingfield MJ, de Beer ZW. (2017) Biodiversity and ecology of flower-associated actinomycetes in different flowering stages of Protea repens. Antonie van Leeuwenhoek 10.1007/s10482-017-0942-3
McTaggart AR, Beasley DR, Wingfield MJ, Wood AR, Pretorius ZA, Drenth A, Shivas RG, Roux J. (2017) A dynamic, web-based resource to identify rust fungi (Pucciniales) in southern Africa. MycoKeys 26:77-83. 10.3897/mycokeys.26.14602
De Maayer P, Aliyu H, Vikram S, Blom J, Duffy B, Cowan DA, Smits THM, Venter SN, Coutinho TA. (2017) Phylogenomic, pan-genomic, pathogenomic and evolutionary genomic insights into the agronomically relevant enterobacteria Pantoea ananatis and Pantoea stewartii. Frontiers in Microbiology 8(1755) 10.3389/fmicb.2017.01755
Jami F, Wingfield MJ, Gryzenhout M, Slippers B. (2017) Diversity of tree-infecting Botryosphaeriales on native and non-native trees in South Africa and Namibia. Australasian Plant Pathology 10.1007/s13313-017-0516-x
Stewart JE, Ross-Davis AL, Graҫa RN, Alfenas AC, Peever TL, Hanna JW, Uchida JY, Hauff RD, Kadooka CY, Kim MS, Cannon PG, Namba S, Simeto S, Pérez CA, Rayamajhi MB, Lodge DJ, Arguedas M, Medel-Ortiz R, López-Ramirez MA, Tennant P, Glen M, Machado PS, McTaggart AR, Carnegie AJ, Klopfenstein NB, Cleary M. (2017) Genetic diversity of the myrtle rust pathogen (Austropuccinia psidii) in the Americas and Hawaii: Global implications for invasive threat assessments. Forest Pathology 10.1111/efp.12378