Development of primary cell cultures from the adult xylem-feeding leafhopper, Kolla paulula, as a tool for studying Wolbachia biology

Xylophagous leafhoppers are vectors of xylem-limited plant pathogens such as citrus variegated chlorosis (CVC) and Pierce's disease, which cause Xylella diseases. Currently, no cure for Xylella diseases exists. The objective of endosymbiont control using Wolbachia pipientis is to reduce the populations of insect vectors, potentially preventing the expansion of Xylella diseases. The purpose of this study was to establish primary cell cultures from adult xylem-feeding leafhoppers to study Wolbachia biology. Cells from adult male and female Kolla paulula (Walker) (Hemiptera: Membracoidea: Cicadellinae) were successfully cultured in Dulbecco's Modified Eagle Medium containing 20% fetal bovine serum and were maintained for more than 6 months. Cells of both male and female adults are round and semi-attached. The doubling times for male and female K. paulula cells are approximately 8 and 10 days, respectively. The presence of Wolbachia in K. paulula cell cultures was detected by polymerase chain reaction (PCR) amplification of Wolbachia surface protein (wsp) gene. The cell cultures developed in this study may be useful in studying interactions between Wolbachia and its hosts of different genders.