Host plant identification in the generalist xylem feeder Philaenus spumarius through gut content analysis

The meadow spittlebug, Philaenus spumarius L. (Hemiptera: Aphrophoridae), is the main vector of the phytopathogenic bacterium Xylella fastidiosa in Europe, where the ST53 strain induces the olive quick decline syndrome, causing severe economic damage in southern Italy. The wide range of plant species infected by X. fastidiosa, and the wide host range of P. spumarius suggest that a large number of wild and cultivated plants may become infected by the pathogen following unintentional introduction events. Therefore, it is necessary to detail the host plant preference of the vector, in order to include preferred plants in the field, in pathogen-targeted diagnostic efforts. This would allow the identification of main sources of X. fastidiosa acquisition by P. spumarius; such plant species represent an important target for rational disease management. Here, we investigated the host plants of P. spumarius in north-western Italy, a region where X. fastidiosa is still not present but is regarded as a primary threat. We designed a new molecular diagnostic tool targeting chloroplast DNA, to characterize the gut content of single P. spumarius adults. The newly set up, nested PCR/sequencing-based identification protocol was proven to be useful for retrieving sequences from the last two different host plants used by P. spumarius, even if limited persistence of intact chloroplast DNA was reported in the spittlebug gut. We propose this protocol as a new tool for supporting research on xylem feeder biology that could be particularly useful for highly polyphagous species such as P. spumarius. Furthermore, the method could help monitor X. fastidiosa invasion, and contribute to the study of vector ecology and pathogen epidemiology.