The ultrastructure of mature spermatozoa of three cicada species, Subpsaltria
yangi, Karenia
caelatata, and Platypleura
kaempferi, was investigated using epifluorescence and transmission electron microscopies. This is the first investigation of the sperm ultrastructure of species in the subfamily Tibicininae and the tribe Sinosenini, represented by S.
yangi and K.
caelatata, respectively. The three species all produce two or three types of spermatozoa with various lengths, viz., polymegaly. The centriolar adjunct of spermatozoa in S.
yangi shows a granular substructure, which is different from that of other cicada species, suggesting that spermatozoa in Tibicininae may have their own characteristics in comparison with other cicadas. The centriolar adjunct of spermatozoa of K.
caelatata displays characteristics similar to that of the Cicadinae. Combined with other morphological characters, it is reasonable to remove K.
caelatata and its allies (i.e., Sinosenini) from Cicadettinae to Cicadinae. The study of sperm ultrastructure, particularly in the species of Tibicininae and Sinosenini, expands the spermatological research of Cicadidae and provides more information for phylogenetic analysis of Cicadidae.