qPCR Assays for Sensitive and Rapid Detection of Quambalaria Species from Plant Tissues

Several species from the genus (order Microstromatales) cause diseases on eucalypts ( and related genera) both in plantations and natural ecosystems. We developed real-time quantitative PCR (qPCR) assays to rapidly detect and distinguish five species. The design of the species-specific qPCR assay for each species, (PIT), (COR), (CYN), (PUS), and (EUC), was based on the ITS region and was evaluated for specificity and sensitivity. The PIT, COR, and CYN qPCR assays could amplify as little as 10 fg µl from pure cultures, whereas PUS and EUC qPCR assays could amplify 100 fg µl of their target species. The PIT, COR, and CYN qPCR assays were further validated using naturally and artificially infected samples of their plant host These assays will be used for rapid diagnostics and future experiments on the infection process.