SYSTEMATICS Phylogenetic Relationships among Subfamilies in Higher Termites (Isoptera: Termitidae) Based on Mitochondrial COII Gene Sequences TORU MIURA, KIYOTO MAEKAWA, OSAMU KITADE, TAKUYA ABE, 1 AND TADAO MATSUMOTO Department of Biology, University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan Ann. Entomol. Soc. Am. 91(5): 515-523 (1998) ABSTRACT Among the 7 families of Isoptera, higher termites (family Termitidae) have acquired the most complex social systems and widest diversity of feeding habits in the course of adaptive radiation. Termitidae is composed of 4 subfamilies (Apicotermitinae, Macrotermitinae, Nasutiter- mitinae, and Termitinae); however, the phylogenetic relationships among subfamilies remains ambiguous. To test the monophyly of this family and the relationship among the 4 subfamilies, we sequenced the mitochondrial cytochrome oxidase II (COII) gene in 15 termite genera of Termitidae and closely related Rhinotermitidae. DNA sequences and translated protein sequences were ana- lyzed using parsimony and the neighbor-joining method. All the resulting trees gave robust support for the monophyly of Termitidae, and overall the relationship of Macrotermitinae + (Apicotermiti- nae + [Termitinae + Nasutitermitinae]) was supported, which is consistent with previous studies based on morphology. KEY WORDS Termitidae, molecular phylogeny, mitchondrial DNA, COII gene, relationships among 4 subfamilies ONE OF THE major groups of eusocial insects is the order Isoptera (termites), which is currently recog- nized as having 7 families and 14 subfamilies (Krishna 1970, Kambhampati et al. 1996). Since the earliest classification of families, termites have been divided into 2 major groups: the lower and higher termites (Snyder 1949, Emerson 1955; reviewed in Krishna 1969). This designation is based mainly on the pres- ence of symbiotic protozoa in the hindgut of lower termites. Higher termites consist of only 1 family, Termitidae, which contains 4 subfamilies—Termiti- nae, Apicotermitinae, Macrotermitinae, and Nasuti- termitinae (Krishna 1970, Sands 1972). Termitidae ex- hibits a wide variety of social specialization and a rigid caste system with highly stereotyped postembryonic development (Krishna 1969, Noirot 1969). Moreover, it is the largest family in Isoptera, containing 3/4 of all known species. So far, =230 genera have been de- scribed in Termitidae (Pearce and Waite 1994). Most of these species are tropical, and abundance is highest in the area close to the equatorial regions (Collins 1983). Until the 1970s, the classification of subfamilies dif- fered from the present one: all genera now in Apico- termitinae, and some in Termitinae, were classified into Amitermitinae. It was thought that Amitermitinae and Termitinae were closely related to each other, and Macrotermitinae and Nasutitermitinae shared certain primitive characters (Emerson 1945, Ahmad 1950; re- 1 Center for Ecological Research, Kyoto University, Shimosaka- moto, Ohtsu, Shiga 520-0105, Japan. viewed in Krishna 1970). Although Grass6 and Noirot (1954) erected a new subfamily, Apicotermitinae, it was not accepted widely. Sands (1972) presented the phylogeny of higher termites based on intestinal and mandible morphology. He stressed that Apicotermiti- nae was an independent subfamily and the rest of Amitermitinae was included in Termitinae. The gut morphology of Nasutitermitinae was clearly derived from Termitinae. Macrotermitinae, with a short intes- tine like those of the more primitive families of the lower termites, was distinct from the other 3 subfam- ilies. Therefore, the inferred relationship was Macro- termitinae + (Apicotermitinae + [Nasutitermitinae + Termitinae]). Phylogenetic relationships among termite families has been reported recently, based on mitochondrial 16S ribosomal RNA gene fragment (Kambhampati et al. 1996). This work showed that Mastotermitidae was the most basal taxon in termites, and Termopsidae was more basal than Kalotermitidae, which was the sister taxon of the Rhinotermitidae-Termitidae complex. In the study, however, the monophyly of Termitidae was not supported strongly. In addition, the relationships among subfamilies in higher termites (Termitidae) has not been examined based on molecular data. In this study we sequenced the mitochondrial cy- tochrome oxidase II (COII) gene and analyzed the phylogenetic relationship using neighbor-joining and most-parsimony method, in order to clarify the mono- phyly of Termitidae and the relationships among the four subfamilies of higher termites. 0013-8746/98/0515-0523302.00/0 © 1998 Entomological Society of America Downloaded from https://academic.oup.com/aesa/article/91/5/515/24057 by guest on 28 September 2021