Mr Donovin Coles
I joined the FABI family in 2015 as part of my honours project. The technical aspect of this project focused on developing a CRISPR-Cas9 construct to knock-out a nitrate transporter 2.5 (NRT2.5) gene, to functionally characterize its role in plant defence. This gene has been implicated during defence against a broad-spectrum of diseases in Eucalyptus grandis. The CRISPR-Cas9 genome editing technology has recently become the preferred tool to functionally characterize genes due to its high specificity and versatility in a wide-range of plant species. NEBuilder® HIFI DNA assembly was used to develop a CRISPR-Cas9 construct to knock-out NRT2.5 in Arabidopsis thaliana (Arabidopsis). Furthermore, a phylogenetic tree was constructed to identify the putative Eucalyptus grandis NRT2.5 orthologs and the starting material for the CRISPR system was designed to target a Eucalyptus grandis NRT2.5 ortholog. This study will contribute to future studies, aimed at elucidating the role of NRT2.5 in plant defence, but more importantly in Eucalyptus defence.
In 2016, I started my MSc: Biotechnology which is a continuation of my honours project under the supervision of Dr Sanushka Naidoo. This project aims to utilize the CRISPR-Cas9 construct developed during my honours year to create functional knock-outs of NRT2.5 in Arabidopsis. These Arabidopsis NRT2.5 knock-out lines will then undergo pathogenicity trials to elucidate the role of this gene in plant defence. Once a role in the model plant Arabidopsis has been elucidated, focus will be driven towards studies in Eucalyptus. This project will hopefully shed some light on defence strategies in Eucalyptus as well as provide CRISPR tools for future functional studies in this economically important hardwood tree.