Transcriptomic characterization of Wolbachia endosymbiont from Leuronota fagarae (Hemiptera: Psylloidae)

<b>Aim:</b> <i>Wolbachia</i> species are among the most abundant intracellular endosymbionts of insects worldwide. The extensive distribution of Gram-negative <i>Wolbachia</i> among insects highlights their evolutionary success and close relationship with many insect host species. This study aimed to characterize a novel <i>Wolbachia</i> strain from the Wild Lime Psyllid, <i>Leuronota fagarae</i> (<i>L. fagarae</i>), to understand its evolutionary relationship with <i>Wolbachia</i> from psyllid pests like <i>Diaphorina citri</i>, the vector of Huanglongbing (HLB). <b>Methods:</b> Wild-caught <i>L. fagarae</i> colonies from Florida, USA, were maintained on Zanthoxylum fagara. RNA was extracted from the salivary glands, heads, and whole bodies of male and female adult <i>L. fagarae</i>. Four cDNA libraries were sequenced using short read technology and de novo transcriptome assembly was performed. Multilocus sequence typing (MLST) of nine conserved loci and <i>wsp</i> gene analysis classified the strain's phylogeny, while sequence mapping and functional annotation provided insight into host-microbe interactions. <b>Results:</b> The new <i>Wolbachia</i> strain, designated <i>Wolbachia</i> endosymbiont of <i>Leuronota fagarae</i> (wLfag-FL), was assigned to supergroup B, showing relation to <i>Wolbachia</i> strains of other related psyllids. Transcriptome analysis identified 1,359 <i>Wolbachia</i> transcripts with 465 assigned functions encompassing metabolic and secretion system pathways. Ankyrin domain proteins and a partial bacterioferritin sequence were detected, suggesting nutritional provisioning roles. <b>Conclusion:</b> The characterization of wLfag-FL expands the known <i>Wolbachia</i> host range and informs HLB-related pest biology. Its phylogenetic placement and transcript annotations offer insights into symbiotic interactions, potentially guiding environmentally safe pest control strategies targeting psyllid fitness and pathogen transmission.