Phyllode inoculation provides a rapid protocol for preliminary screening of Acacia species for tolerance to Ceratocystis wilt and canker disease

Ceratocystis wilt and canker disease has devastated Acacia mangium plantations in south-east Asia. Current screening methodologies to identify resistant or tolerant germplasm use potted plants in a greenhouse as a preliminary screening to select material for field trials, but these tests are time-consuming, motivating a search for rapid screening protocols. In this study, inoculation procedures were tested on three species of Acacia, viz. A. mangium, A. crassicarpa and a hybrid of A. auriculiformis and A. mangium, using three isolates of Ceratocystis manginecans. Mycelial plugs were compared with spore suspensions as inoculum to infect artificial wounds on the stems of A. mangium potted plants. The rapid screening protocols involved inoculation of stem segments with mycelial plugs and detached phyllodes with a spore suspension, with susceptibility measured by lesion length on the stems or necrosis length on the phyllodes. Both mycelial plugs and spore suspensions produced a similar level of disease incidence, so either inoculum form can be used for an assay. The stem segments were prone to contamination by other fungi and to desiccation, while results from the potted plant and phyllode protocols showed similar trends of susceptibility among the Acacia clones and species. The ease, rapidity, and reproducibility of the phyllode inoculation protocol makes it a potential replacement for inoculation of potted plants as a preliminary screening protocol to identify disease tolerant A. mangium germplasm prior to field screening.