Optimisation of sampling and testing for asymptomatic olive trees infected by Xylella fastidiosa in Apulia region, Italy

Early detection of Xylella fastidiosa outbreaks in Apulian olive groves is crucial, especially in buffer zones and pathogen-free areas where olive trees are asymptomatic. Three studies were conducted. Two were on the spatial and tempo-ral progression of X. fastidiosa infections in tree canopies of asymptomatic or mildly symptomatic olive trees of tolerant (`Leccino') and susceptible (`Cellina di Nardo' and `Ogliarola salentina') cultivars. Despite different trends in pathogen infection rates and concentrations between `Leccino' and susceptible olive cultivars over the study period, results showed that sampling was most effective in the mid-upper part of tree cano-pies throughout the year, excluding the warmest and coldest periods. Stem xylem tis-sues were the most appropriate for detecting the pathogen compared to lower parts of mature leaves with petioles, using serological and molecular assays. Based on these results, a third study was conducted to compare molecular and serological tests (qPCR, real-time LAMP, DAS-ELISA, DTBIA) for detection of X. fastidiosa in the mid-upper part of asymptomatic branches of infected `Leccino' trees that were sam-pled in an appropriate collection time, using stem xylem tissue as the most appropri-ate matrix for testing. The molecular methods showed the greatest sensitivity, with no undetermined results, while among the serological assays, DTBIA was more sensitive than DAS-ELISA. An improved protocol for monitoring asymptomatic olive trees is recommended.