Continuous cell lines derived from the Asian citrus psyllid, Diaphorina citri, harbor viruses and Wolbachia

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is a major pest of global citriculture. In the Americas and in Asia, D. citri vectors the phloem-limited bacterium, Candidatus Liberibacter asiaticus (CLas), which causes the fatal citrus disease huanglongbing, or citrus greening. Cell lines derived from D. citri can provide insight into both the basic biology of this pest and D. citri-associated pathogens including CLas. We previously identified CLG#2 as the optimal medium for long-term growth of D. citri primary cell cultures. Here we report on the establishment and characterization of three continuous D. citri cell lines, Dici1, Dici3, and Dici5, that have been passaged for > 40 times. Based on morphological and transcriptomic data, the Dici1 and Dici3 cell lines include undifferentiated and neurogenic progenitor cells. Dici1 and Dici5 are infected with Wolbachia. Both Dici1 and Dici5 are infected with D. citri reovirus, and Dici5 is also infected with D. citri-associated C virus. Dici3 is free of both Wolbachia and virus infection. These cell lines provide an ideal platform for the study of inter-microbial relationships as well as microbe interaction with host insect cells.