Transferring an Agrobacterium-mediated transformation protocol across eight genera in the Ceratocystidaceae

Abstract

Establishing a transformation system is an essential first step for functional studies. Numerous transformation systems have been developed and optimized for filamentous ascomycetes, including Agrobacterium-mediated transformation. Recently, such a system was developed for the African tree pathogen Ceratocystis albifundus. This fungus is part of the family Ceratocystidaceae, which includes many tree and plant pathogens of economic importance. Despite the many advantages associated with Agrobacterium-mediated transformation, establishing and optimizing this system within a species are extremely arduous due to the many species-specific parameters that must be optimized. This study aimed to apply the existing C. albifundus Agrobacterium-mediated transformation protocol to eight diverse Ceratocystidaceae species from eight different genera. Minor adjustments were made to the already established protocol to increase transformant yield before it was used to successfully produce transformants for six of the eight targeted species without the need for any further species-specific optimizations. Although the number of transformants obtained was less than for C. albifundus, this study proves that protocols for Agrobacterium-mediated transformation might be more transferable than previously thought. It also provides a useful benchmark for future transformation studies in the Ceratocystidaceae and will make species-specific optimization easier.