Identification of three potential insect vectors of Xylella fastidiosa in southern Italy

In order to identify potential vectors of Xylella fastidiosa in olive orchards in Puglia (southern Italy), Hemiptera insects were collected from October to December, 2013, in olive orchards with high incidences of X. fastidiosa associated with "rapid decline" symptoms. The study focused on species in the Auchenorrhyncha (sharpshooter leafhoppers and froghoppers or spittlebugs), a group that includes known vectors of X. fastidiosa. Adults of three species, i.e. Philaenus spumarius L. (Aphrophoridae), Neophilaenus campestris Fallen (Aphrophoridae) and Euscelis lineolatus Brulle (Cicadellidae) were captured, from which total DNA was extracted and assayed by PCR using three sets of specific primers designed for X. fastidiosa detection. Results of PCR showed that 38 out of a total of 84 tested insects were positive for X. fastidiosa, i.e. eight (of 20) P. spumarius, 14 (of 18) N. campestris and 16 (of 46) E. lineolatus. PCR amplicons of the RNA polymerase sigma-70 factor gene from six specimens (two of each insect species) were sequenced. The sequences obtained were 99.3-99.4% identical. BlastN analyses demonstrated these sequences to be similar to those of X. fastidiosa isolates from olive OL-X and OL-G reported from Puglia, whereas they displayed distant molecular identity (always less than 98%) with X. fastidiosa subspecies from other countries. The detection of X. fastidiosa in P. spumarius and, for the first time, in N. campestris and E. lineolatus (which, unlike the others, is a phloem feeder), indicates potential vectoring roles of these insects for the spread of the bacterium in Puglia. Further investigations and specific infectivity trials are required to definitively determine the roles of these insects as effective vectors of this pathogen.