Optimising Agrobacterium‑mediated transformation of Corynespora cassiicola to deliver DsRed

Murnita B Mohmad Mahyudin, Gary D Foster, Andrew M. Bailey

Research output: Contribution to journalArticle (Academic Journal)peer-review

Abstract

Corynespora cassiicola is a cosmopolitan fungus with a broad host range. One of the hosts often severely impacted is the rubber
tree, Hevea brasiliensis on which it causes devastating leaf defoliation. To date, control of the disease is mainly focused
on the application of fungicides, but this only provides short-term solution. Functional genomics through transformation
allows evaluation of the disease effectors which may eventually lead to more precise control measures to curb the disease.
A transformation system for C. cassiicola mediated by Agrobacterium tumefaciens was successfully established with resistance
to hygromycin and DsRed as markers. The protocol was optimised with regard to Agrobacterium strains, C. cassiicola
isolates, and other parameters. Combination of isolate CLN23 and A. tumefaciens strain LBA4404 carrying pCAMDsRed
gave the highest number of transformants. The optimum fungal age, temperature for incubation and co-cultivation period
were 7 days old, 28 °C and 4 days, respectively. However, these parameters are highly subjected to interaction between the
variables. The transformants carried the DsRed reporter gene, and demonstrated readily apparent red fluorescence, which
allowed disease development in planta to be observed. This robust transformation system should pave the way for future
genetic manipulations of C. cassiicola.
Original languageEnglish
Pages (from-to)139-149
JournalJournal of Rubber Research
Volume23
DOIs
Publication statusPublished - 6 Jun 2020

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