Wolbachia strain w Lhui induces temperature‐dependent incomplete cytoplasmic incompatibility in the invasive pest Liriomyza huidobrensis with biocontrol potential

BACKGROUND Wolbachia is a maternally inherited endosymbiont that manipulates host reproduction through cytoplasmic incompatibility (CI), offering promising opportunities for biocontrol of agricultural pests. The leaf‐miner Liriomyza huidobrensis (Blanchard) is a globally invasive and highly polyphagous pest with a high incidence of Wolbachia infection; however, its reproductive effects remain poorly understood. Here, we investigated the reproductive manipulation induced by the Wolbachia strain w Lhui using genomics analyses and crossing assays. RESULTS w Lhui localized primarily to the reproductive tissues of both female and male adults and maintained a 100% infection prevalence across three host generations under both low (15 and 20 °C) and moderate (25 °C) temperatures. Crossing assays showed that w Lhui induced incomplete CI, reducing egg hatch by approximately 30% in incompatible crosses. Both CI strength ( s h ) and w Lhui density varied with host rearing temperature. Genome sequencing revealed that w Lhui (approximately 1.27 Mb) belongs to supergroup A and harbors two pairs of CI factor genes ( cifA and cifB ). These Cif proteins are classified as Type I and exhibit substantial phylogenetic and structural divergence. Expression of CifB‐pair1 caused growth defects in yeast, suggesting that CifB‐pair1 exhibits toxicity. However, no direct interaction between CifA and CifB was detected by yeast two‐hybrid assays. CONCLUSIONS These findings elucidate the role and molecular basis of w Lhui‐induced reproductive manipulation and highlight its potential for developing Wolbachia ‐based biocontrol strategies against leaf‐miner pests. © 2026 Society of Chemical Industry.