Liriomyza chinensis (Kato) is a formidable pest of Allium species, especially the Japanese bunching onion Allium fistulosum L. Recently, a novel biotype of L. chinensis (biotype B) has emerged, which causes more severe damage than the native biotype A. It has been reported that biotype B has frequently displaced biotype A in the Japanese bunching onion fields in Japan. As interbiotype hybridization is a possible factor that influences such displacement, interbiotype hybridization was conducted between L. chinensis biotypes A and B. Eggs were not laid under one-by-one crossing conditions; however, adult hybrid progeny of both sexes emerged from no-choice mating combinations-when multiple males and females were present. The fertility of F1 hybrid adults was also investigated, and backcrossed adults emerged from F1 females in both mating combinations. F1 males might have exhibited reproductive abnormalities because only a small number of backcross progeny emerged from the mating combinations using F1 males. Additionally, 3 representative endosymbionts (Wolbachia, Spiroplasma, and Cardinium) were investigated, and both biotypes were found to be infected by the same strain of Wolbachia. In addition, the courtship signals (tapping) of male adults differed between biotypes A and B as well as between F1 hybrids; the F1 males exhibited tapping behavior that was intermediate between biotypes A and B. Therefore, mating sounds serve as a form of premating reproductive isolation between biotypes A and B.© The Author(s) 2023. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.