CRISPR/Cas9-mediated genome editing of gustatory receptor NlugGr23a causes male sterility in the brown planthopper Nilaparvata lugens.

Gustatory receptors (Grs) have an essential role in chemical recognition so as to evaluate food quality. Insect Grs also participate in non-gustatory functions, such as olfaction, temperature sensing, and mating. In this study, we knocked out NlugGr23a, a putative fecundity-related Gr, using the CRISPR/Cas9 system in the brown planthopper Nilaparvata lugens, a serious insect pest of rice. Surprisingly, homozygous NlugGr23a mutant (NlugGr23a-/-) males were sterile but their sperm were motile and morphologically normal. DAPI staining of mutant sperm inseminated eggs showed that most of NlugGr23a-/- sperm failed to fertilize eggs, even if they were capable of entering into the egg as a result of their arrested development prior to male pronucleus formation. Immunohistochemistry demonstrated the expression of NlugGr23a in testis. Moreover, prior mating by NlugGr23a-/- males suppressed female fertility. To our knowledge, it is the first report that a chemoreceptor is implicated in male sterility and provides a potential molecular target for genetic pest control alternatives.